KR20240065302A - Combination therapy for cancer using biphenyl compounds and immune checkpoint molecular modulators - Google Patents

Abstract

A novel cancer treatment method is provided that exhibits remarkably excellent anti-tumor effects with few side effects. Antitumor agents include biphenyl compounds for use in the treatment of subjects with cancer. Treatment includes administering an immune checkpoint molecule modulator to the subject.

Description

Combination therapy for cancer using biphenyl compounds and immune checkpoint molecular modulators

The present disclosure relates to an anti-tumor agent comprising a biphenyl compound or a salt thereof in combination with an immune checkpoint molecule modulator; And it relates to the manufacture of anti-tumor effect enhancing agents and kits.

Lysine-specific demethylase 1 (LSD1) is an epigenetic enzyme that mono- or dimethylates histone H3 lysine 4 (H3K4) and H3 lysine 9 (H3K9). In addition to its function as an epigenetic regulator, LSD1 regulates some non-histone substrates, including DNMT1, p53, STAT3, and E2F1 (Non-Patent Document 1). LSD1 is also a component of different multi-protein complexes, and its association with various gene regulatory proteins has been demonstrated (Non-patent Document 2). Numerous studies have highlighted the central role of LSD1 in the control of several cellular processes in normal and cancer cells, such as stemness, differentiation, cell motility, and epithelial-to-mesenchymal transition (Non-Patent Document 3).

LSD1 is highly expressed in many cancers and is associated with poor prognosis (Non-Patent Document 4). It is involved in various stages of cancer, including development, progression, metastasis, and recurrence after therapy. Targeting LSD1 has been recognized as a promising strategy for cancer treatment in recent years. Several LSD1 inhibitors are undergoing clinical evaluation for cancer therapy (Non-Patent Document 1).

Meanwhile, the immune system is an important mechanism for self-defense against various diseases caused by factors in vivo and in vitro. Deterioration of the function of the immune system has detrimental pathological effects, such as the development of infectious diseases caused by bacteria and viruses, the development of tumors, and a delay in recovery from injuries and diseases. Therefore, activation of the immune system is very important for the prevention and treatment of various diseases. As one of the new methods for treating cancer, cancer immunotherapy is being developed.

Activation of the adaptive immunological response is initiated by binding of the antigenic peptide-MHC complex to the T-cell receptor (TCR). Additionally, this binding is regulated by co-stimulation or co-inhibition due to binding between the B7 family of costimulatory molecules and the CD28 family of receptors of the B7 family. That is, two characteristic signaling events are required for T-cells to be activated in an antigen-specific manner, and T-cells that have not undergone co-stimulation from the B7 family and have only undergone antigen-stimulation are in an unresponsive state (unresponsive). is converted to induce immunological tolerance in T-cells.

By utilizing this mechanism, cancer cells suppress the activation of antigen-specific T-cells and continue to grow outside of immune surveillance. Therefore, for cancer treatment, it is considered effective to control immune evasion of the tumor by inducing an in vivo anti-tumor immune response in cancer patients by enhancing costimulation and blocking costimulation, and controlling immune evasion of the tumor by promoting co-stimulation and blocking co-inhibition. Various cancer immunotherapies have been proposed targeting co-inhibitory molecules (inhibitory co-stimulatory molecules) or co-inhibitory molecules (inhibitory co-stimulatory molecules) (Patent Document 6). For example, as an immune checkpoint molecule modulator to activate T-cells by inhibiting the binding of PD-1 and its ligands (PD-L1 and PD-L2), nivolumab (human-type against human PD-1) IgG4 monoclonal antibody) is used for malignant melanoma, etc. (Patent Document 1 and Non-Patent Document 5).

Biphenyl compound 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-( 2-Hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof is known as an LSD1 inhibitor (Patent Document 2), and is a combination of an LSD1 inhibitor and another antitumor agent. This has been reported so far (Patent Document 3).

However, the biphenyl compound represented by formula (I) and the immune checkpoint molecule modulator have not yet been used in combination. Additionally, the immunostimulatory action of the biphenyl compound represented by formula (I) is not known.

List of cited references

patent literature

PTL 1: International Publication No. WO2004/004771

PTL 2: International Publication No. WO2017/090756

PTL 3: International Publication No. WO2018/216795

non-patent literature

NPL 1: J. Hematol. Oncol, 12: 129 (2019)

NPL 2: Cancers (Basel), 11: 324 (2019)

NPL 3: Crit. Rev. Eukaryot. Gene Expr, 22: 53-59 (2012)

NPL 4: Pharmacol. Res, 164: 105335 (2020)

NPL 5: N. Engl. J. Med., 366: 2443-2454 (2012)

The purpose of the present disclosure is to provide a novel cancer treatment method that exhibits significantly excellent anti-tumor effects with few side effects.

4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]- The combination of 2-fluoro-benzonitrile (“Compound A”) or a salt thereof and an immune checkpoint molecular modulator was studied, and the anti-tumor effect of the combination was investigated. As a result, it was found that the anti-tumor effect was significantly enhanced without causing serious side effects compared to when a single agent was used.

Specifically, in one embodiment, the present disclosure provides the following aspects [1] to [26].

[1] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-) for use in the treatment of subjects with cancer An antitumor agent comprising hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof, wherein the treatment comprises administering an immune checkpoint molecule modulator to the subject. Antitumor agent.

[2] The anti-tumor agent according to [1], wherein the immune checkpoint molecular modulator is at least one selected from the group consisting of a PD-1 pathway antagonist and a CTLA-4 pathway antagonist.

[3] The antitumor agent according to [1] or [2], wherein the immune checkpoint molecule modulator is a PD-1 pathway antagonist.

[4] The anti-tumor agent of [2] or [3], wherein the PD-1 pathway antagonist is at least one selected from the group consisting of anti-PD-1 antibody, anti-PD-L1 antibody, and anti-PD-L2 antibody. my.

[5] The anti-tumor agent according to [2] or [3], wherein the PD-1 pathway antagonist is an anti-PD-1 antibody or an anti-PD-L1 antibody.

[6] The anti-tumor agent according to [2] or [3], wherein the PD-1 pathway antagonist is an anti-PD-1 antibody.

[7] The anti-tumor agent according to [2], wherein the CTLA-4 pathway antagonist is an anti-CTLA-4 antibody.

[8] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl) as active ingredient ) phenyl] phenyl] -2-fluoro-benzonitrile or a salt thereof, an anti-tumor effect enhancing agent for immune checkpoint molecular modulators.

[9] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl] An antitumor agent for treating cancer patients given an immune checkpoint molecule modulator, comprising phenyl]-2-fluoro-benzonitrile or a salt thereof.

[10] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy), containing immune checkpoint molecular modulators -2-Methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof is given as an antitumor agent for treating cancer patients.

[11] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2- for use in treating tumors by combined administration with immune checkpoint molecule modulators Fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof.

[12] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro- for use in enhancing the antitumor effect of immune checkpoint molecular modulators. 4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof.

[13] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro for use in treating tumors in cancer patients given immune checkpoint molecular modulators. Ro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof.

[14] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl] An immune checkpoint molecule modulator for use in treating tumors in cancer patients given phenyl]-2-fluoro-benzonitrile or a salt thereof.

[15] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy- A combination of 2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and an immune checkpoint molecular modulator.

[16] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro for preparing antineoplastic agents administered in combination with immune checkpoint molecular modulators. Use of -4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof.

[17] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[ for preparing an anti-tumor effect enhancer to enhance the anti-tumor effect of immune checkpoint molecular modulators. Use of 2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof.

[18] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2 for preparing antineoplastic agents for treating cancer patients given immune checkpoint molecular modulators. Use of -fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof.

[19] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl] Use of an immune checkpoint molecule modulator for the manufacture of an anti-tumor agent for the treatment of cancer patients given phenyl]-2-fluoro-benzonitrile or a salt thereof.

[20] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2) for producing antitumor agents -Methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or salts thereof and use of immune checkpoint molecular modulators.

[21] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy) to a subject in need of treatment for a tumor A method of treating a tumor comprising administering a combination of -2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and an immune checkpoint molecule modulator.

[22] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluorotherapy for subjects in need of enhancement of the anti-tumor effect of immune checkpoint molecular modulators. A method for enhancing the antitumor effect of an immune checkpoint molecular modulator, comprising administering -4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. .

[23] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy) to a subject in need of treatment for a tumor A method of treating a tumor in a cancer patient given an immune checkpoint molecule modulator, comprising administering -2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof.

[24] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[, comprising administering an immune checkpoint molecule modulator to a subject in need of treatment for a tumor. A method of treating tumors in a cancer patient given 2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof.

[25] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy) to a subject in need of treatment for a tumor A method of treating a tumor comprising administering a combination of -2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and an immune checkpoint molecular modulator.

[26] 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl] A pharmaceutical composition for preventing and/or treating tumors, comprising phenyl]-2-fluoro-benzonitrile or a salt thereof and an immune checkpoint molecule modulator.

Additionally, another exemplary embodiment relates to:

- 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl, administered in combination )phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and an antitumor agent comprising an immune checkpoint molecule modulator.

- 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl- as monotherapy or combination therapy A method of treating cancer patients exhibiting higher levels of gMDSC using an antitumor agent comprising propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and/or an immune checkpoint molecule modulator.

- 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2 in cancer patients based on the amount of granulocytic myeloid-derived suppressor cells (gMDSC) in samples isolated from cancer patients. -[2-Fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and/or an antitumor agent comprising an immune checkpoint molecule modulator A method of treating cancer, comprising predicting chemotherapy using and administering an anti-tumor agent to any cancer patient predicted to respond sufficiently to chemotherapy using an anti-tumor agent.

- 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2 in cancer patients based on the amount of granulocytic myeloid-derived suppressor cells (gMDSC) in samples isolated from cancer patients. -[2-Fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and/or an antitumor agent comprising an immune checkpoint molecule modulator A method for predicting chemotherapy for cancer treatment, comprising predicting chemotherapy using.

The anti-tumor agent of the present disclosure makes it possible to perform cancer treatment with high anti-tumor effect while suppressing the occurrence of side effects.

[Figure 1A] Figure 1A shows the ratio of granulocytic myeloid-derived suppressor cells (gMDSC, same as polymorphonuclear-MDSC) in tumor infiltrating leukocytes.
[Figure 1B] Figure 1B shows the ratio of CD8 positive T cells to tumor infiltrating leukocytes.
[Figure 2a] Figure 2a shows the anti-tumor effect of Compound A in a wild-type (immune competent) mouse model transplanted with the mouse colon cancer cell line MC38.
[FIG. 2B] FIG. 2B shows the anti-tumor effect of Compound A in an immunodeficient mouse model transplanted with the mouse colon cancer cell line MC38.
Figure 3 Figure 3 depicts the effect of the combination of Compound A and anti-mouse PD-1 antibody in a wild-type mouse model transplanted with the mouse colon cancer cell line MC38.

4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]- 2-Fluoro-benzonitrile is shown in the following structure:

In this disclosure, this compound is described as “Compound A”. Compound A is described as Example Compound 37 in PCT Publication No. WO2017/090756, the disclosure of which is incorporated herein by reference in its entirety. Compound A may be prepared by any method known in the art, including but not limited to those described in PCT Publication Nos. WO2017/090756 and WO2021/095835, the disclosures of which are incorporated herein by reference in their entirety. It is included as.

The present disclosure provides an anti-tumor agent (interchangeable with “medication for treating a tumor”), comprising Compound A or a salt thereof and an immune checkpoint molecule modulator (particularly an anti-PD-1 antibody) administered in combination. may be used), anti-tumor effect enhancers and kit preparations; his uses; How to treat a tumor; and methods for enhancing anti-tumor effects.

Compound A or its salt used in the present disclosure may be in the form of crystals. Single crystals and polymorphic crystal mixtures are included within the scope of Compound A or salts thereof. These crystals can be produced by crystallization according to crystallization methods known in the art. Compound A or a salt thereof may be a solvate (e.g., a hydrate) or a non-solvate. Any such form is included within the scope of the compound or salt thereof of the present disclosure.

Compound A labeled with isotopes (e.g., ³ H, ¹⁴ C, ³⁵ S and ¹²⁵ I) is also included within the scope of Compound A or salts thereof as used in this disclosure.

As used in the present disclosure, salts of compound A refer to common salts used in the field of organic chemistry. Examples of such salts include base addition salts and acid addition salts. The salt of Compound A is preferably a pharmaceutically acceptable salt.

Examples of base addition salts include alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as calcium salts and magnesium salts; ammonium salt; and organic amine salts such as trimethylamine salt, triethylamine salt, dicyclohexylamine salt, ethanolamine salt, diethanolamine salt, triethanolamine salt, procaine salt, and N,N'-dibenzylethylenediamine salt. Includes.

Examples of acid addition salts include inorganic acid salts such as hydrochloride, sulfate, nitrate, phosphate and perchlorate; Organic acid salts such as acetate, formate, maleate, fumarate, tartrate, citrate, ascorbate, benzoate and trifluoroacetate; and sulfonates such as methanesulfonate, isethionate, benzenesulfonate, and p-toluenesulfonate.

One example of a salt of Compound A is the benzoic acid salt or benzoate salt. Another example of a salt of Compound A is sorbic acid, succinic acid, L-tartaric acid, or hydrochloric acid. Another example of a salt of Compound A is hemi-fumarate, mono-oxalate, mesylate, esylate, maleate, mono-fumartate, or hemi-oxalate.

Immune checkpoint molecule modulators in the present disclosure act directly on immune checkpoint molecules to induce an anti-tumor immune response in cancer patients in vivo, thereby controlling immune evasion of tumors.

Examples of immune checkpoint molecular modulators include substances that promote the function of costimulatory molecules (stimulatory costimulatory molecules) or substances that inhibit the function of co-inhibitory molecules (inhibitory costimulatory molecules). Examples of immune checkpoint molecules include the B7 family (B7-1, B7-2, PD-L1, PD-L2, etc.), CD28 family (CTLA-4, PD-1, etc.), TNF superfamily (4-1BBL and OX40L) ), TNF receptor superfamily (4-1BB and OX40) molecules, TIGIT pathway molecules (TIGIT, etc.), and LAG-3 pathway molecules (LAG-3, etc.). For immune checkpoint molecule modulators, agents targeting one or more types of immune checkpoint molecules can be used. Examples of substances include PD-1 pathway antagonists, ICOS pathway agonists, CTLA-4 pathway antagonists, CD28 pathway agonists, BTLA pathway antagonists, 4-1BB pathway agonists, TIGIT pathway antagonists, and LAG-3 pathway antagonists.

In the present disclosure, the immune checkpoint molecular modulator is preferably at least one selected from the group consisting of a PD-1 pathway antagonist, an ICOS pathway agonist, a CTLA-4 pathway antagonist, and a CD28 pathway agonist, from the viewpoint of suppressing side effects. Preferably, it is at least one selected from the group consisting of PD-1 pathway antagonist and CTLA-4 pathway antagonist, and even more preferably, it is PD-1 pathway antagonist.

PD-1 pathway antagonists inhibit immunosuppressive signals from PD-1 expressed on T-cells. PD-L1 or PD-L2 is a ligand of PD-1, and examples of PD-1 pathway antagonists include anti-PD-1 antibody, anti-PD-L1 antibody, anti-PD-L2 antibody, PD-1 extracellular domain , PD-L1 extracellular domain, PD-L2 extracellular domain, PD-1-Ig (fusion protein of PD-1 extracellular domain and FC region of immunoglobulin (Ig)), PD-L1-Ig, PD- Includes L2-Ig, PD-1 siRNA, PD-L1 siRNA and PD-L2 siRNA. The PD-1 pathway antagonist is preferably an anti-PD-1 antibody, an anti-PD-L1 antibody or an anti-PD-L2 antibody, more preferably an anti-PD-1 antibody or an anti-PD-L1 antibody, particularly preferred. It is an anti-PD-1 antibody.

CTLA-4 pathway antagonists inhibit immunosuppressive signals from CTLA-4 expressed on T-cells. B7-1 (CD80) or B7-2 (CD86) are ligands of CTLA-4. The CTLA-4 pathway antagonist is preferably an anti-CTLA-4 antibody, CTLA-4 extracellular domain, CTLA-4-Ig, anti-B7-1 (CD80) antibody or anti-B7-2 (CD86) antibody, Preferably it is an anti-CTLA-4 antibody or CTLA-4-Ig, particularly preferably an anti-CTLA-4 antibody.

In one aspect of the disclosure, the immune checkpoint molecular modulator is preferably at least one selected from the group consisting of anti-PD-1 antibody, anti-PD-L1 antibody and anti-CTLA-4 antibody, more preferably Preferably, it is at least one antibody selected from the group consisting of anti-PD-1 antibody and anti-PD-L1 antibody, particularly preferably anti-PD-1 antibody.

Examples of antibodies include immunoglobulins (IgA, IgD, IgE, IgG, IgM, IgY, etc.), Fab fragments, F(ab') ₂ fragments, single-stranded antibody fragments (scFv), single domain antibodies, and diabodies. (Reference [Nat. Rev. Immunol., 6:343-357, 2006]). These antibodies include monoclonal or polyclonal antibodies, such as human antibodies, humanized antibodies, chimeric antibodies, mouse antibodies, llama antibodies and chicken antibodies.

Humanized IgG monoclonal antibodies or human IgG monoclonal antibodies are preferred.

In the present disclosure, anti-PD-1 antibodies include nivolumab, pembrolizumab, zimberelimab, cemiplimab, spartalizumab, budigallimab, camrelizumab, cetrelimab, dostarlimab, ezaben. Rimab, lambrolizumab, pempulimab, pimivalimab, fucotenlimab, sasanlimab, sintilimab, ticelizumab, toripalimab, etc., preferably nivolumab, pembrolizumab or zimberelimab.

In the present disclosure, the anti-PD-L1 antibody is atezolizumab, durvalumab, avelumab, garibulimab, rodapolimab, cosibelimab, socazolimab, etc., preferably atezolizumab, durvalumab or Avelumab, more preferably atezolizumab.

In the present disclosure, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, quavonlimumab, zaliprelimumab, etc., preferably ipilimumab.

In the present disclosure, CTLA-4-Ig is abatacept, etc., preferably abatacept.

Anti-TIGIT antibodies in the present disclosure include vivostolimab, osiferlimab, dombanalimab, tiragolumab, and the like.

Anti-LAG-3 antibodies in the present disclosure include relatlimab, pavezelimab, pianlimab, and the like.

These antibodies can be produced by conventional methods for producing antibodies. Moreover, commercially available antibodies can also be used.

In the present disclosure, when two or more immune checkpoint molecular modulators are used, for example, an anti-PD-1 antibody and an anti-CTLA-4 antibody or an anti-PD-1 antibody and an anti-LAG-3 antibody. They can be used in combination, or bispecific antibodies that can bind to both PD-1 and CTLA-4 or both PD-1 and LAG-3 can be used. Examples of bispecific antibodies include XmAb20717 (PD-1 x CTLA-4) and tebotelimab (PD-1 x LAG-3).

In the present disclosure, the daily dose of Compound A or a salt thereof is preferably the recommended single dose of Compound A or a salt thereof from the viewpoint of the enhancing effect on the anti-tumor effect of the immune checkpoint molecule modulator by Compound A. It is 50 to 200%, more preferably 75 to 150%, and especially preferably 100%.

A typical daily dose of Compound A or a salt thereof may range from 100 picograms to 100 milligrams per kilogram of body weight, more typically from 10 nanograms to 25 milligrams per kilogram of body weight. More typically, the daily dose of Compound A or a salt thereof may range from 100 nanograms to 20 milligrams per kilogram of body weight, although higher or lower doses may be administered as needed. For example, the daily dose may be 1 microgram to 20 milligrams per kilogram of body weight, more typically 10 micrograms to 20 milligrams per kilogram of body weight, and more typically 100 micrograms to 20 milligrams per kilogram of body weight.

Dose can also be expressed as the amount of drug administered relative to the patient's body surface area (mg/m ² ). A typical daily dose of Compound A or a salt thereof may range from 3700 pg/m ² to 3700 mg/m ² , but higher or lower doses may be administered if necessary. For example, the daily dose may be 370 ng/m ² to 925 mg/m ² , more typically 3700 ng/m ² to 740 mg/m ² , although higher or lower doses may be administered as needed. there is. For example, 37 micrograms/m ² to 740 mg/m ² , and more typically 370 micrograms/m ² to 740 mg/m ² or 3700 micrograms/m ² to 740 mg/m ² .

Compound A of the present disclosure or its salt can be administered orally in a single dose range, for example, 0.05 to 3000 mg. Typically, the range may be 10 to 1000 mg. Typical examples of capacities are 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900 and 1000. Contains mg. The dosage may be increased or decreased in a stepwise manner from any of the above ranges (0.05 to 3000 mg), for example in increments/decrements of 1 mg, 5 mg, 10 mg, 20 mg, or 50 mg.

In the present disclosure, the daily dose of the immune checkpoint molecular modulator is preferably the recommended dose of the immune checkpoint molecular modulator administered as a single dose, in view of the enhancing effect on the anti-tumor effect of the immune checkpoint molecular modulator by Compound A. Preferably it is 50 to 200%, more preferably 75 to 150%, and especially preferably 100%.

Typical examples of recommended doses of nivolumab administered as a single dose are 2 mg/kg (body weight) per dose or 3 mg/kg (body weight) per dose.

A typical example of a recommended dose of pembrolizumab administered as a single dose is 2 mg/kg (body weight) per dose or 200 mg per dose.

A typical example of a recommended dose of single administered atezolizumab is 1200 mg per dose.

A typical example of a recommended dosage of single administered avelumab or durvalumab is 10 mg/kg (body weight) per dose.

A typical example of a recommended dose of single administered ipilimumab is 3 mg/kg (body weight) per dose.

In the present disclosure, the “recommended dosage” is determined through clinical trials, etc., and is the dosage that produces the maximum therapeutic effect while ensuring safe use without the occurrence of serious side effects. Specifically, the “recommended dosage” refers to public authorities or companies, such as the Pharmaceuticals and Medical Devices Agency (PMDA), the Food and Drug Administration (FDA) and the European Medicines Agency. ) (EMA), the dosage approved, recommended or proposed and described in the accompanying literature, interview forms, treatment guidelines, etc., preferably approved by a public authority selected from the group consisting of PMDA, FDA and EMA.

The administration schedule of the anti-tumor agent of the present disclosure can be appropriately selected depending on the type of cancer, stage of disease, etc.

The administration schedule of Compound A or its salt is not particularly limited as long as it includes a continuous administration period of 1 or 2 weeks, followed by a rest period of 1 week. A cycle is defined as one (1) cycle when a 2-week (14-day) dosing schedule consisting of 1 week (7 days) of continuous dosing followed by a 1-week (7-day) rest period is defined as one (1) cycle. It may be performed once or repeated two or more times. That is, administration may be performed in a specific administration schedule of one cycle or more than one cycle, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 cycles or more. In some embodiments, administration can be carried out over an extended period of time, including multiple cycles. For example, administration may be approximately 13 to 15 cycles of treatment or administration over a 6-month period; With approximately 25 or 26 cycles of treatment over a 1-year period; Approximately 75 to 100 cycles of treatment or more may be performed over a 3-year period. If a 3-week (21-day) dosing schedule consisting of 2 weeks (14 days) of dosing followed by a 1-week (7-day) rest period is defined as one (1) cycle, the cycle may be performed once or repeated two or more times. It can be. That is, administration may be performed in a dosing schedule of one cycle or more than one cycle, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 cycles or more. In some cases, administration may be performed in more cycles. For example, administration may be approximately 10 cycles over a period of 6 months; 1-year period with approximately 20 cycles; It can be performed over a 3-year period with approximately 50 to 60 cycles or a longer dosing period with more cycles.

Additionally, in the dosing schedule for Compound A, as long as the dosing continues on a schedule of 1 or 2 weeks of continuous dosing followed by a 1 week washout period, the dosing may be discontinued thereafter, and the dosing may be continued after a specific washout period (no dosing). It can be resumed. Similarly, the dosing schedule of the present disclosure may include a schedule with multiple drug wash periods. In one embodiment of a dosing schedule with a washout period, it is sufficient that the condition “1 or 2 weeks of continuous administration followed by a washout period of 1 week” is met in the dosing period before the washout period and in the dosing period after the washout period. In another embodiment of a dosing schedule with two washout periods, the condition “1 or 2 weeks of continuous dosing followed by a 1 week washout period” is the dosing period before the first washout period, the dosing period between the two washout periods. and in the administration period after the second drug-free period. In another embodiment of a dosing schedule with two or more washout periods, the condition “1 or 2 weeks of continuous dosing followed by a 1 week washout period” is a dosing period preceding the first washout period, between two adjacent washout periods. It is sufficient to meet the requirements during the administration period and the administration period following the last drug holiday period. The drug holiday period is not particularly limited and can be set appropriately depending on the patient's condition, etc. For example, the drug holiday period may range from 1 to 35 days. Alternatively, the drug holiday period may range from 1 to 12 months.

The administration schedule of the immune checkpoint molecular modulator is also not particularly limited as long as it is administered at specific intervals. In some embodiments, the immune checkpoint molecule modulator is administered at intervals of 1 to 3 weeks.

The administration schedule of nivolumab is preferably one in which administration is performed at 2 or 3 week intervals.

The dosing schedule for pembrolizumab, atezolizumab or ipilimumab is preferably one in which administration is performed at three-week intervals.

The administration schedule of avelumab or durvalumab is preferably one in which administration is performed at two-week intervals.

The number of daily administrations of the anti-tumor agent of the present disclosure is appropriately selected depending on the type of cancer, stage of the disease, etc.

The number of times Compound A or its salt is administered per day is preferably 1 or 2 times, more preferably 1 time. The number of doses of nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab or ipilimumab per day is preferably once.

The order of administration of Compound A or its salt and the immune checkpoint molecular modulator in the present disclosure may be appropriately selected depending on the type of cancer, stage of the disease, etc., and these agents are used in any order or in combination. Can be administered in parallel.

“In combination” can refer to the use of one agent in the treatment of a subject, wherein the treatment includes administering another agent to the subject.

Here, when Compound A or its salt and the immune checkpoint molecule modulator are not administered in parallel, the administration interval between the two agents can be appropriately selected as long as the enhancing effect on the antitumor effect is exhibited, and the administration interval is preferable. Preferably it is 1 to 14 days, more preferably 1 to 7 days, even more preferably 1 to 5 days, especially preferably 1 to 3 days.

The tumor targeted in the present disclosure is not particularly limited as long as it exhibits an enhancing effect on the anti-tumor effect, and the tumor is preferably a tumor in which Compound A or a salt thereof exhibits an anti-tumor effect, and more preferably a malignancy involving LSD1. It's a tumor.

The type of malignant tumor treated by Compound A or a salt thereof is not particularly limited. Examples of such malignancies include glandular tumors, carcinoid tumors, undifferentiated carcinomas, angiosarcomas, adenocarcinomas, gastrointestinal cancers (e.g., colorectal cancer (“CRC”), e.g., colon and rectal cancers, biliary tract cancers, e.g. Gallbladder and bile duct cancers (cholangiocarcinomas), anal cancers, esophageal cancers, gastrointestinal (stomach) cancers, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (“GISTs”), liver cancers, duodenal cancers, and small intestine cancers), lung cancers (e.g. Cellular lung cancer (“NSCLC”), squamous-cell lung carcinoma, large cell lung carcinoma, small cell lung carcinoma, invasive mucinous adenocarcinoma, mesothelioma and other lung cancers such as bronchial tumors and pleuropulmonary carcinomas), urinary cancers (e.g., kidney Cancer (renal cancer), transitional cell carcinoma of the kidney (“TCC”), TCC of the renal pelvis and ureter (“PDQ”), bladder cancer, urethral cancer, and prostate cancer), head and neck cancer (e.g., eye cancer, retinoblastoma, intraocular melanoma) Tumor, hypopharyngeal cancer, pharynx cancer, laryngeal cancer, laryngeal papillomatosis, metastatic squamous cervical cancer as an occult primary, sinonasal squamous cell carcinoma (SNSCC), oral cancer (oral cancer), lip cancer, pharynx cancer, oropharyngeal cancer, sensory neuroblastoma, nasal cancer. and paranasal cancer, nasopharyngeal cancer, and salivary gland cancer), endocrine cancer (e.g., thyroid cancer, parathyroid cancer, multiple endocrine neoplasia syndrome, thymoma and thymic carcinoma, pancreatic cancer (e.g., pancreatic ductal adenocarcinoma (“PDAC”), pancreatic cancer) neuroendocrine tumors and islet cell tumors), breast cancer (extrahepatic ductal carcinoma in situ (“DCIS”), lobular carcinoma in situ (“LCIS”), triple negative breast cancer, and inflammatory breast cancer), male and female reproductive cancers (e.g. Cervical cancer, ovarian cancer, endometrial cancer, uterine sarcoma, uterine cancer, vaginal cancer, vulvar cancer, gestational trophoblast tumor (“GTD”), extragonadal germ cell tumor, extracranial germ cell tumor, germ cell tumor, testicular cancer, and penile cancer) , brain and nervous system cancers (e.g., astrocytoma, brainstem glioma, brain tumor, glioblastoma (GBM), craniopharyngioma, central nervous system (“CNS”) cancer, chordoma, ependymoma, embryonal tumor, neuroblastoma , paraganglioma and atypical malformation, oligodendroma, oligodendroastrocytoma, oligodendroglioma, anaplastic oligodendroastrocytoma, ganglioglioma, central neurocytoma, medulloblastoma, germ cell tumor, meningioma, schwannoma, GH secretion. pituitary adenoma, PRL-secreting pituitary adenoma, ACTH-secreting pituitary adenoma, dysfunctional pituitary adenoma, hemangioblastoma, and epidermoid tumor), skin cancer (e.g., basal cell carcinoma (“BCC”), squamous cell skin carcinoma (“ SCC"), Merkel cell carcinoma and melanoma), tissue and bone cancers (e.g., soft-tissue sarcoma, rhabdomyosarcoma, fibrous histiocytoma of bone, Ewing's sarcoma, malignant fibrous histiocytoma of bone ("MFH") , osteosarcoma and chondrosarcoma), cardiovascular cancer (e.g., heart cancer and cardiac tumors), appendix cancer, childhood and adolescent cancer (e.g., adrenocortical carcinoma of childhood, midline ductal carcinoma, hepatocellular carcinoma (“HCC”) , hepatoblastoma and Wilms carcinoma) and virus-induced cancers (e.g., HHV-8 associated cancer (Kaposi's sarcoma) and HIV/AIDS associated cancer).

Tumors also suitable for treatment will include hematological and plasma cell malignancies (e.g., cancers affecting the blood, bone marrow and/or lymph nodes) such as multiple myeloma, leukemia and lymphoma, myelodysplastic syndromes and myeloproliferative disorders. However, it is not limited to this. Leukemias include, but are not limited to, acute lymphoblastic leukemia (“ALL”), acute myeloid (“AML”) leukemia, chronic lymphocytic leukemia (“CLL”), chronic myeloid leukemia (“CML”), acute monocytic leukemia (“AMoL”), hairy cell leukemia, and/or other leukemias. Lymphomas include, but are not limited to, Hodgkin's lymphoma and non-Hodgkin's lymphoma (“NHL”). In some embodiments, the NHL is B-cell lymphoma and/or T-cell lymphoma. In some embodiments, NHL includes, but is not limited to, diffuse large B-cell lymphoma (“DLBCL”), small lymphocytic lymphoma (“SLL”), chronic lymphocytic leukemia (“CLL”), mantle cell lymphoma (“MCL”). "), Burkitt's lymphoma, cutaneous T-cell lymphomas, including mycosis fungoides and Sézary syndrome, AIDS-related lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma (Waldenstrom's macroglobulinemia ("WM")), primary central nervous system (CNS) lymphoma, central nervous system malignant lymphoma, and/or other lymphoma.

More preferably, examples include lung cancer (non-small cell lung cancer, small cell lung cancer, etc.) and leukemia. More preferably, examples include small cell lung cancer (SCLC) and acute myeloid leukemia (AML).

As shown in the examples below, Compound A or its salt has an immunostimulatory effect. As used herein, “immunostimulatory action” refers to the action of activating immune cells, such as cytotoxic T cells and macrophages, and/or suppressing the inhibition of immune responses by regulatory T cells, myeloid-derived suppressor cells (MDSC), etc. means. In particular, Compound A or a salt thereof has the effect of increasing cytotoxic T cells and/or suppressing the suppression of immune responses by granulocytic myeloid-derived suppressor cells (gMDSC).

In the present disclosure, gMDSCs can be identified by markers commonly known to those skilled in the art. Examples of markers are CD33 ⁺ CD11b ⁺ HLA-DR ^- CD15 ⁺ CD14 ^- , CD33 ⁺ CD11b ⁺ HLA-DR ^low CD15 ⁺ CD14 ^- , CD33 ⁺ CD11b ⁺ Lin ^- CD15 ⁺ CD14 ^- , CD33 ⁺ CD11b ⁺ HLA-DR ^- Lin ^- CD15 ⁺ CD14 ^- and CD33 ⁺ CD11b ⁺ HLA-DR ^low Lin ^- CD15 ⁺ CD14 ^- .

In the present disclosure, the active ingredient Compound A or a salt thereof and the immune checkpoint molecule modulator may be separated and formulated into multiple dosage forms or integrated and formulated into one dosage form depending on the formulation or administration schedule of the active ingredient. (i.e., formulated as a combination preparation). The formulations may be manufactured and sold in one package suitable for combined administration or may be manufactured and sold in individual packages.

The formulation of the anti-tumor agent according to the present disclosure is not particularly limited and may be appropriately selected depending on the purpose of treatment, and specific examples thereof include oral formulations (tablets, coated tablets, powders, granules, capsules, solutions, etc.), injections, etc. , suppositories, patches and ointments.

In the case of Compound A or its salt, oral formulations are preferred.

In the case of anti-PD-1 antibody, anti-PD-L1 antibody or anti-CTLA-4 antibody, the above-mentioned agents are used, and injection is preferred.

For both Compound A or a salt thereof and an immune checkpoint molecular modulator, the anti-tumor agent according to the present disclosure will typically be prepared by known methods using one or more pharmaceutically acceptable carriers depending on the formulation of the agent. You can. Examples of carriers include various carriers commonly used in common pharmaceutical agents, such as excipients, binders, disintegrants, lubricants, diluents, solubilizers, suspending agents, isotonic agents, pH adjusters, buffers, stabilizers, colorants, flavors. Includes improvers and odor improvers.

The anti-tumor agent of the present disclosure makes it possible to perform cancer treatment with high anti-tumor effect while suppressing the occurrence of side effects. In some embodiments, the high anti-tumor effect includes a tumor growth delay effect, a tumor reduction effect, or both. Therefore, when an anti-tumor agent is administered to a cancer patient, the patient's lifespan is extended.

The present disclosure also relates to an anti-tumor effect enhancing agent for enhancing the anti-tumor effect of an immune checkpoint molecular modulator on cancer patients, wherein the anti-tumor effect enhancing agent contains Compound A or a salt thereof as an active ingredient. The anti-tumor effect enhancing agent has the same formulation form as the above-mentioned anti-tumor agent.

The present disclosure also relates to an anti-tumor effect enhancing agent containing an immune checkpoint molecule modulator as an active ingredient for enhancing the anti-tumor effect of Compound A or a salt thereof on cancer patients. The anti-tumor effect enhancing agent has the same formulation form as the above-mentioned anti-tumor agent.

The present disclosure also relates to an anti-tumor agent for treating cancer patients given an immune checkpoint molecule modulator containing Compound A or a salt thereof. Antitumor agents have the above-mentioned formulation forms.

The present disclosure also relates to anti-tumor agents containing immune checkpoint molecule modulators for treating cancer patients given Compound A or a salt thereof. Antitumor agents have the above-mentioned formulation forms.

“Treatment” includes postoperative adjuvant chemotherapy performed to prevent recurrence after surgical removal of the tumor and preoperative adjuvant chemotherapy performed before surgical removal of the tumor.

The present disclosure also relates to an anti-tumor agent containing Compound A or a salt thereof used in combination with an immune checkpoint molecule modulator for cancer patients. Antitumor agents have the above-mentioned formulation forms.

The present disclosure also relates to an anti-tumor agent containing an immune checkpoint molecule modulator, used in combination with an anti-tumor agent containing Compound A or a salt thereof for cancer patients. Antitumor agents have the above-mentioned formulation forms.

The present disclosure also relates to anti-tumor agents containing Compound A or a salt thereof; and written instructions indicating that Compound A or a salt thereof and an immune checkpoint molecule modulator are administered in combination to a cancer patient.

The "written instructions" herein are not limited as long as the above-mentioned dosages are specified. Written instructions may or may not be legally binding and preferably the above-mentioned dosages are recommended. Specific examples of written guidance include attached documents and brochures. Kit preparations containing written instructions may have written instructions printed or affixed to the kit manufacturing package, or the written instructions may be included with the anti-tumor agent within the kit manufacturing package.

The present disclosure can be further combined with one or more anti-cancer agents other than the anti-tumor agents described above. Examples of anticancer agents include chemotherapy agents (eg, cytotoxic agents), immunotherapy agents, hormonal and antihormonal agents, targeted therapy agents, and antiangiogenic agents. Many anticancer drugs can be classified within one or more of these groups. Although certain anticancer agents are grouped within specific group(s) or subgroup(s) herein, many of these agents are also grouped within one or more other group(s) or subgroup(s) as currently understood in the art. can be listed. Anticancer agents are not particularly limited, and examples thereof include chemotherapy agents, mitotic inhibitors, plant alkaloids, alkylating agents, antimetabolites, platinum analogs, enzymes, topoisomerase inhibitors, retinoids, aziridines, antibiotics, hormones, and antihormones. agents, anti-estrogens, anti-androgens, anti-adrenal agents, androgens, targeted therapy agents, immunotherapy agents, biological response modulators, cytokine inhibitors, tumor vaccines, monoclonal antibodies, colony-stimulating factors, anti-LAGl agonists, anti-OX40 Including, but not limited to, agents.

Non-limiting examples of chemotherapeutic agents include mitotic inhibitors, plant alkaloids, alkylating agents, antimetabolites, platinum analogs, enzymes, topoisomerase inhibitors, retinoids, aziridines, and antibiotics.

Non-limiting examples of mitotic inhibitors and plant alkaloids include taxanes such as cabazitaxel, docetaxel, larotaxel, ortataxel, paclitaxel, and tesetaxel; demecolcine; epothilone; eribulin; etoposide (VP-16); etoposide phosphate; Navelbin; noscapine; teniposide; taliblastin; Vinblastine; vincristine; vindesine; vinflunine; and vinorelbine.

Non-limiting examples of alkylating agents include nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, cytophosphan, estramustine, ifosfamide, mannomustine, mechlorethamine, mechlorethamine oxide. Hydrochloride, melphalan, nobombikine, phenesterine, prednimustine, tris(2-chloroethyl)amine, trophosphamide and uracil mustard; Alkyl sulfonates such as busulfan, improsulfan and fiposulfan; Nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine, streptozotocin, and TA-07; ethyleneimine and methylamelamine, such as altretamine, thiotepa, triethylenemelamine, triethylenethiophosphaoramide, triethylenephosphoramide and trimethylolomelamine; Ambamustine; bendamustine; Dacarbazine; cyclophosphamide; etoglucide; Irofulven; mafosfamide; mitobronitol; mitolactol; pipobroman; procarbazine; temozolomide; Treosulfan; and triaziquone.

Non-limiting examples of antimetabolites include folic acid analogs such as aminopterin, denopterin, edatrexate, methotrexate, pteropterin, raltitrexed, and trimetrexate; Purine analogs such as 6-mercaptopurine, 6-thioguanine, fludarabine, porodecine, thiamiprine and thioguanine; Pyrimidine analogs such as 5-fluorouracil (5-FU), tegafur/gimeracil/oteracil potassium, tegafur/uracil, trifluridine, trifluridine/tipiracil hydrochloride, 6- Azauridine, ancitabine, azacitidine, capecitabine, carmofur, cytarabine, decitabine, dideoxyuridine, doxypiuridine, doxyfluridine, enocitabine, floxuridine, galocitabine. , gemcitabine and safacitabine; 3-aminopyridine-2-carboxaldehyde thiosemicarbazone; Broxuridine; cladribine; cyclophosphamide; cytarabine; Emitepur; hydroxyurea; mercaptopurine; Nellarabine; pemetrexed; pentostatin; Tegapur; and troxacitabine.

Non-limiting examples of platinum analogs include carboplatin, cisplatin, dicycloplatin, heptaplatin, lobaplatin, nedaplatin, oxaliplatin, satraplatin, and triplatin tetranitrate.

Non-limiting examples of enzymes include asparaginase and pegaspargase.

Non-limiting examples of topoisomerase inhibitors include acridine carboxamide, amonapide, amsacrine, belotecan, elliptinium acetate, exatecan, indolocarbazole, irinotecan, lurtotecan, mitoxantrone, Includes joxane, rubitecan, SN-38, soxane, and topotecan.

Non-limiting examples of retinoids include alitretinoin, bexarotene, fenretinide, isotretinoin, liarosole, RII retinamide, and tretinoin.

Non-limiting examples of aziridines include benzodopa, carboquone, meturedopa, and uredopa.

Non-limiting examples of antibiotics include implantable antibiotics; anthracenedione; Anthracycline antibiotics such as aclarubicin, amrubicin, daunomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, menogaril, nogalamycin, pirarubicin and valrubicin; 6-diazo-5-oxo-L-norleucine; aclasinomycin; actinomycin; Outhramycin; Azaserine; bleomycin; Cactinomycin; Calicheamicin; carabicin; carminomycin; Carzinophilin; Chromomycin; dactinomycin; Detorubicin; esorubicin; esperamicin; geldanamycin; Marcellomycin; mitomycin; Mitomycin C; mycophenolic acid; Olivomycin; Novantrone; peplomycin; porphyromycin; Portiromycin; puromycin; kellamycin; rebeccamycin; rhodolubicin; streptonigrin; streptozocin; Thanespimycin; tubersidin; Ubenimex; Ginostatin; Ginostatin stimalamer; and zorubicin.

Non-limiting examples of hormonal and antihormonal agents include antiandrogens such as abiraterone, apalutamide, bicalutamide, darolutamide, enzalutamide, flutamide, goserelin, leuprolide, and nilutamide; Antiestrogens, such as 4-hydroxy tamoxifen, aromatase inhibitor 4(5)-imidazole, EM-800, fospestrol, fulvestrant, keoxifene, LY 117018, onapristone, raloxifene, tamoxifen, toremifene and trioxyphene; Anti-adrenal agents such as aminoglutethimide, dexaminoglutethimide, mitotane, and trilostane; Androgens such as calusterone, dromostanolone propionate, epithiostanol, mephithiostane and testolactone; abarelix; Anastrozole; cetrorelix; deslorelin; exemestane; Fadrozole; finasteride; formestane; histrelin (RL 0903); human chorionic gonadotropin; lanreotide; LDI 200 (Milkhaus); Letrozole; leuprorelin; mifepristone; Nafarelin; Nafoxidine; Osaterone; prednisone; thyrotropin alpha; and triptorelin.

Non-limiting examples of immunotherapeutic agents (i.e., immunotherapy) include biological response modifiers, cytokine inhibitors, tumor vaccines, monoclonal antibodies, colony-stimulating factors, and immunomodulators.

Non-limiting examples of biological response modifiers, including cytokine inhibitors (cytokines) such as interferons and interleukins, include interferon alpha/interferon alpha, such as interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-nl, interferon alpha- n3, interferon alpha interferon-1, peginterferon alpha-2a, peginterferon alpha-2b and leukocyte alpha interferon; Interferon beta, such as interferon beta-1a and interferon beta-1b; interferon gamma, such as natural interferon gamma-1a and interferon gamma-1b; aldesleukin; interleukin-1 beta; interleukin-2; Ofrelbekin; sonermin; tasonermin; and virulizine.

Non-limiting examples of tumor vaccines include APC 8015, AVICINE, bladder cancer vaccine, cancer vaccine (Biomira), gastrin 17 immunogen, Maruyama vaccine, melanoma lysate vaccine, melanoma tumor lysate vaccine (New York New York Medical College), Melanoma Vaccine (New York Medical College), Melanoma Vaccine (Sloan Kettering Institute), TICE(R) BCG (Bacillus Calmette-Guerin) ), and viral melanoma cell lysate vaccine (Royal Newcastle Hospital).

Non-limiting examples of monoclonal antibodies include abagovomab, adecatumumab, aflibercept, alemtuzumab, blinatumomab, brentuximab vedotin, CA 125 MAb (Biomira), cancer MAb (Japan Pharma) Japan Pharmaceutical Development), daclizumab, daratumumab, denosumab, edrecolomab, gemtuzumab zogamicin, HER-2 and Fc MAb (Medarex), Ibri Tumomab tiuxetan, idiotype 105AD7 MAb (CRC Technology), idiotype CEA MAb (Trilex), lintuzumab, LYM-1-iodine 131 MAb (Techni clone) ), mitumomab, moxetumomab, ofatumumab, polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), ranibizumab, rituximab, veltuzumab, and trastuzumab.

Non-limiting examples of colony-stimulating factors include darbepoetin alfa, epoetin alfa, epoetin beta, filgrastim, granulocyte macrophage colony stimulating factor, lenograstim, leridistim, myrimostim, molgramostim, nar Includes tograstim, pegfilgrastim, and sargramostim.

Non-limiting examples of additional immunotherapeutic agents include BiTE, CAR-T cells, GITR agonists, imiquimod, immunomodulatory imide (IMiD), mismatched double-stranded RNA (ampligen), resiquimod, SRL. 172, and thymalfacin.

Targeted therapies include, for example, monoclonal antibodies and small molecule drugs. Non-limiting examples of targeted therapies include signal transduction inhibitors, growth factor inhibitors, tyrosine kinase inhibitors, EGFR inhibitors, HER2 inhibitors, histone deacetylase (HDAC) inhibitors, proteasome inhibitors, cell-cycle inhibitors, angiogenesis inhibitors, matrix -Metalloproteinase (MMP) inhibitors, hepatocyte growth factor inhibitors, TOR inhibitors, KDR inhibitors, VEGF inhibitors, fibroblast growth factor (FGF) inhibitors, RAF inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, AKT inhibitors, MCL -1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, BRAF-inhibitors, RAS inhibitors, gene expression modulators, autophagy inhibitors, apoptosis inducers, antiproliferative agents, and glycolysis inhibitors.

Non-limiting examples of signal transduction inhibitors include tyrosine kinase inhibitors, multi-kinase inhibitors, alotinib, avapritinib, axitinib, dasatinib, dovitinib, imatinib, lenvatinib, ronidamine, nilotinib, nintedanib. , pazopanib, pegvisomant, ponatinib, vandetanib, and EGFR and/or HER2 inhibitors.

Non-limiting examples of EGFR inhibitors include small molecule antagonists of EGFR such as afatinib, brigatinib, erlotinib, gefitinib, lapatinib, neratinib, dacomitinib, vandetanib, and osimertinib; and antibody-based EGFR inhibitors, such as any anti-EGFR antibody or antibody fragment that can partially or completely block EGFR activation by its natural ligand. Antibody-based EGFR inhibitors are described, for example, in Modjtahedi, H., et al., 1993, Br. J. Cancer 67:247-253; Teramoto, T., et al., 1996, Cancer 77:639-645; Goldstein et al., 1995, Clin. Cancer Res. 1:1311-1318; Huang, S. M., et al., 1999, Cancer Res. 15:59(8): 1935-40; and Yang, X., et al., 1999, Cancer Res. 59: 1236-1243; monoclonal antibody Mab E7.6.3 (Yang, 1999, supra); Mab C225 (ATCC Accession No. HB-8508), or an antibody or antibody fragment with its binding specificity; specific antisense nucleotides or siRNA; Afatinib, cetuximab; matuzumab; necitumumab; Nimotuzumab; Panitumumab; and zalutumumab.

Non-limiting examples of HER2 inhibitors include HER2 tyrosine kinase inhibitors such as afatinib, lapatinib, neratinib, and tucatinib; and anti-HER2 antibodies or drug conjugates thereof, such as trastuzumab, trastuzumab emtansine (T-DM1), pertuzumab, margetuximab, trastuzumab deruxtecan (DS-8201a), and trastuzumab Contains zumab duocarmazine.

Non-limiting examples of histone deacetylase (HDAC) inhibitors include belinostat, panobinostat, romidepsin, and vorinostat.

Non-limiting examples of proteasome inhibitors include bortezomib, carfilzomib, ixazomib, marizomib (salinosporamide a), and ofrozomib.

Non-limiting examples of cell-cycle inhibitors, including CDK inhibitors, include abemaciclib, albocidib, palbociclib, and ribociclib.

Non-limiting examples of antiangiogenic agents (or angiogenesis inhibitors) include matrix-metalloproteinase (MMP) inhibitors; VEGF inhibitor; EGFR inhibitor; TOR inhibitors such as everolimus and temsirolimus; PDGFR kinase inhibitors such as crenolanib; HIF-1α inhibitors such as PX 478; HIF-2α inhibitors such as velzutipan and the HIF-2α inhibitors described in WO 2015/035223; Fibroblast growth factor (FGF) or FGFR inhibitors such as B-FGF and RG 13577; Hepatocyte growth factor inhibitor; KDR inhibitor; anti-Ang1 and anti-Ang2 agonists; anti-Tie2 kinase inhibitor; Tek antagonist (US 2003/0162712; US 6,413,932); anti-TWEAK agent (US 6,727,225); ADAM disintegrin domain, which antagonizes the binding of integrins to their ligands (US 2002/0042368); anti-eph receptor and/or anti-ephrin antibody or antigen binding region (US 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447; and 6,057,124); and anti-PDGF-BB antagonists, as well as antibodies or antigen binding regions that specifically bind PDGF-BB ligands.

Non-limiting examples of matrix-metalloproteinase (MMP) inhibitors include MMP-2 (matrix-metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, prinomastat, Includes RO 32-3555 and RS 13-0830. Examples of useful matrix metalloproteinase inhibitors are for example WO 96/33172, WO 96/27583, EP 1004578, WO 98/07697, WO 98/03516, WO 98/34918, WO 98/34915, WO 98/ 33768, WO 98/30566, EP 0606046, EP 0931788, WO 90/05719, WO 99/52910, WO 99/52889, WO 99/29667, WO 1999/007675, EP 1786785, EP 1181017, US 2009/0012085, US 5,863,949, US 5,861,510, and EP 0780386. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity to inhibit MMP-1. Other matrix-metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12 , and MMP-13), it is more preferable to selectively inhibit MMP-2 and/or MMP-9.

Non-limiting examples of VEGF and VEGFR inhibitors include bevacizumab, cediranib, CEP 7055, CP 547632, KRN 633, orantinib, pazopanib, pegaptanib, pegaptanib octansodium, semaxanib, sorafenib, sunitinib. , VEGF antagonist (Borean, Denmark), and VEGF-TRAP ^™ .

Other antiangiogenic agents include, but are not limited to, 2-methoxyestradiol, AE 941, alemtuzumab, alpha-D148 Mab (Amgen, USA), alphastatin, anecortave acetate, angiosidin, angiogenesis. Inhibitors (SUGEN, USA), angiostatin, anti-Vn Mab (Crucell, Netherlands), atiprimod, axitinib, AZD 9935, BAY RES 2690 (Bayer, Germany), BC 1 (Genoa Institute of Cancer Research, Italy), beloranib, Benepin (Lane Labs, USA), cabozantinib, CDP 791 (Celltech Group, UK) ), chondroitinase AC, cilengitide, combretastatin A4 prodrug, CP 564959 (OSI, USA), CV247, CYC 381 (Harvard University, USA), E 7820, EHT 0101, endostatin , enzastaurin hydrochloride, ER-68203-00 (IVAX, USA), fibrinogen-E fragment, Flk-1 (ImClone Systems, USA), form of FLT 1 (VEGFR 1), FR-111142, GCS-100, GW 2286 (GlaxoSmithKline, UK), IL-8, ilomastat, IM-862, irsogladin, KM-2550 (Kyowa Hakko, Japan), lenalidomide, lenvatinib, MAb alpha5beta3 integrin, second generation (Applied Molecular Evolution, USA and MedImmune, USA), MAb VEGF (Xenova) ), UK), marimastat, maspin (Sosei, Japan), metastatin, motuporamine C, M-PGA, ombrabulin, OXI4503, PI 88, platelet factor 4, PPI 2458, Lamu Sirumab, rBPI 21 and BPI-derived antiangiogenesis (XOMA, USA), regorafenib, SC-236, SD-7784 (Pfizer, USA), SDX 103 (University of California, San Diego, USA) , SG 292 (Telios, USA), SU-0879 (Pfizer, USA), TAN-1120, TBC-1635, tesevatinib, tetrathiomolybdate, thalidomide, thrombospondin 1 inhibitor, Tie-2 ligand (Regeneron, USA), tissue factor pathway inhibitor (EntreMed, USA), tumor necrosis factor-alpha inhibitor, tumstatin, TZ 93, urokinase plasminogen activator inhibitor , including vadimezan, vandetanib, vasostatin, vatalanib, VE-cadherin-2 antagonist, xanthorizole, XL 784 (Exelixis, USA), zib-aflibercept, and ZD 6126. can do.

Anticancer agent(s) that may be combined with Compound A may also be activators that destroy or inhibit the RAS-RAF-ERK or PI3K-AKT-TOR signaling pathways. Examples include RAF inhibitors, EGFR inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, MCL-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasome inhibitors or monoclonal antibodies, immunomodulators, etc. Immunotherapies including, but not limited to, IMiDs, anti-LAGl and anti-OX40 agents, GITR agonists, CAR-T cells, and BiTEs.

Non-limiting examples of RAF inhibitors include dabrafenib, encorafenib, regorafenib, sorafenib, and vemurafenib.

Non-limiting examples of MEK inhibitors include binimetinib, CI-1040, cobimetinib, PD318088, PD325901, PD334581, PD98059, lefametinib, selumetinib, and trametinib.

Non-limiting examples of ERK inhibitors include LY3214996, LTT462, MK-8353, SCH772984, raboxertinib, ulixertinib, and ASTX029.

Non-limiting examples of PI3K inhibitors include 17-hydroxywortmannin analogs (e.g., WO 06/044453); AEZS-136; alpelisib; AS-252424; Buparlisib; CAL263; Copanlisip; CUDC-907; Dactolisib (WO 06/122806); demethoxyviridine; duvelisib; GNE-477; GSK1059615; IC87114; idelariship; INK1117; LY294002; Palomid 529; Paxalisib; perifosine; PI-103; PI-103 hydrochloride; pictilisib (e.g., WO 09/036,082; WO 09/055,730); PIK 90; PWT33597; SF1126; sonolisip; TGI 00-115; TGX-221; XL147; XL-765; wortmannin; Taselisib (GDC-0032); and ZSTK474.

Non-limiting examples of AKT inhibitors include Akt-1-1 (inhibits Aktl) (Barnett et al. (2005) Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2 (Barnett et al. (2005) Biochem. J. 385 (Pt. 2), 399-408); API-59CJ-Ome (e.g., Jin et al. (2004) Br. J. Cancer 91, 1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g. WO05011700); indole-3-carbinol and its derivatives (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li (2004) J Nutr. 134(12 Suppl), 3493S-3498S); Perifosine, described in Dasmahapatra et al. (2004) Clin. Cancer Res. 10(15), 5242-52, 2004]); phosphatidylinositol ether lipid analogs (e.g., Gills and Dennis (2004) Expert. Opin. Investig. Drugs 13, 787-97); triciribine (Yang et al. (2004) Cancer Res. 64, 4394-9); trans-3-amino-1-methyl-3-[4-(3-phenyl-5H-imidazo[1,2-c]pyrido[3,4-e][1,3]oxazine-2- 1) imidazooxazone compounds, including phenyl]-cyclobutanol hydrochloride (WO 2012/137870); Apuresertip; Capyvasertip; 8-[4-(1-aminocyclobutyl)phenyl]-9-phenyl-1,2,4-triazolo[3,4-f][1,6]naphthyridin-3(2H)-one (MK2206 ) and pharmaceutically acceptable salts thereof; AZD5363; trans-3-amino-1-methyl-3-(4-(3-phenyl-5H-imidazo[1,2-c]pyrido[3,4-e][1,3]oxazine-2- mono)phenyl)cyclobutanol (TAS117) and its pharmaceutically acceptable salts; and patasertip.

Non-limiting examples of TOR inhibitors include deforolimus; ATP-competitive TORC1/TORC2 inhibitors such as PI-103, PP242, PP30 and Torin 1; TOR inhibitors at the FKBP12 enhancer, rapamycin and its derivatives such as temsirolimus, everolimus, WO 9409010; Rapalogs, e.g. as disclosed in WO 98/02441 and WO 01/14387, e.g. AP23573, AP23464, or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin; 40-epi-(tetrazolyl)-rapamycin (also called ABT578); AZD8055; 32-deoxoraphamycin; 16-fentinyloxy-32(S)-dihydrorapamycin, and other derivatives disclosed in WO 05/005434; US 5,258,389, WO 94/090101, WO 92/05179, US 5,118,677, US 5,118,678, US 5,100,883, US 5,151,413, US 5,120,842, WO 93/111130, WO 94/02136, O 94/02485, WO 95/14023, WO 94 /02136, WO 95/16691, WO 96/41807, WO 96/41807 and US 5,256,790; and phosphorus-containing rapamycin derivatives (eg, WO 05/016252).

Non-limiting examples of MCL-1 inhibitors include AMG-176, MIK665, and S63845.

Non-limiting examples of SHP2 inhibitors include JAB-3068, RMC-4630, TNO155, SHP-099, RMC-4550, and SHP2 inhibitors described in WO 2019/167000, WO 2020/022323, and WO 2021/033153.

Non-limiting examples of RAS inhibitors include AMG510 (sotorasib), MRTX849, LY3499446, JNJ-74699157 (ARS-3248), ARS-1620, ARS-853, RM-007, and RM-008.

Additional non-limiting examples of anti-cancer agents that may be suitable for use include 2-ethylhydrazide, 2,2',2"-trichlorotriethylamine, ABVD, aceglatone, acemannan, aldophosphamide glycoside, Alpharadin, amifostine, aminolevulinic acid, anagrelide, ANCER, Ancestim, anti-CD22 immunotoxin, anti-tumorigenic herb, apaziquone, arglavine, arsenic trioxide, azathioprine, BAM 002 (Novelos), bcl-2 (Genta), Vestrabucil, Viricodar, Bisantrene, Bromocriptine, Brostalysin, Bryostatin, Butionine Sulfoximine, Caliculin, Cell-cycle non-specific antineoplastic agents, selmoreukin, clodronate, clotrimazole, cytarabine oxophosphate, DA 3030 (Dong-A), defopamine, denileukin deftitox, dexrazoxane, Diaziku Onion, dichloroacetic acid, dirazep, discodermolide, docosanol, doxercalciferol, edelfosine, eflornithine, EL532 (Elan), elpomitin, elsamitrucine, enyluracil, etanida Sol, exisulind, feruginol, folic acid supplements such as prolinic acid, achytocin, gallium nitrate, gimeracil/oteracil/tegafur combination (S-1), glycopine, histamine dihydrochloride, HIT diclofenac , HLA-B7 gene therapy (Vical), human fetal alpha fetoprotein, ibandronate, ibandronic acid, ICE chemotherapy regimen, imexone, iobenguan, IT-101 (CRLX101), raniquidar, LC 9018 (Yakult), leflunomide, lentinan, levamisole plus fluorouracil, lovastatin, lucanthone, masoprocol, melarsoprol, metoclopramide, miltefosine, miproxifen. , mitoguazone, mitozolomide, furidamole, motexapine gadolinium, MX6 (Galderma), naloxone plus pentazocine, nitracrine, nolatrexed, NSC 631570 octreotide (Ukrain), Olaparib, P-30 protein, PAC-1, palifermin, pamidronate, pamidronic acid, pentosan polysulfate sodium, phenamet, ficivanil, pixantrone, platinum, podophyllic acid, porfimer sodium, PSK (polysaccharide-K), rabbit anti-thymocyte polyclonal antibody, Raspuri embodiment, retinoic acid, rhenium Re 186 etidronate, romultide, samarium (153 Sm) lexidronam, sizopyran, Sodium phenylacetate, sparphosic acid, spirogermanium, strontium-89 chloride, suramin, swainsonine, talaporphine, tariquidar, tazarotene, tegafur-iodine, tenofurpurine, tethiofurfura. Sil and tipiraparzamine, TLC ELL-12, tositumomab-iodine 131, trifluridine and tipiracil combination, troponin I (Harvard University, USA), urethane, valspodar, verteporfin, zoled. Including, but not limited to, ronic acid, and suzucquidar.

Example

The present disclosure will now be described in further detail by way of examples, which should not be construed as limiting the disclosure, and those skilled in the art will be able to make many modifications within the technical concept of the disclosure. You can.

Example 1: Immunostimulatory action of biphenyl compounds in colon cancer cell transplantation model

The mouse colon cancer cell line MC38 was obtained from Dr. Yoshihiro Hayakawa (University of Toyama, Japan). The MC38 cell strain was cultured in RPMI 1640 medium containing 10% fetal bovine serum and then subcultured at a frequency of once or twice a week in the presence of 5% CO2 in an incubator at 37°C.

MC38 cell suspension was implanted subcutaneously at 1.5 x 10 ⁶ cells/0.1 ml into the right thorax of 6-week-old C57BL/6JJcl mice (CLEA Japan, Inc.).

Tumors were grown until they reached a tumor volume (TV) of 80 to 230 mm ³ after implantation. Digimatic calipers were used to measure tumor diameter. The major and minor diameters of the tumor were measured, and TV was calculated from the equation below.

TV (mm ³ ) = Major diameter (mm) x Minor diameter (mm) x Minor diameter (mm)/2

Five animals were assigned to each group by stratified random assignment using TV as an indicator. The day on which grouping (n=5) was performed was defined as day 0.

To adjust the hypromellose concentration to 0.5 w/v%, an appropriate amount of water for injection as described in the Japanese Pharmacopoeia was added, and then the mixture was stirred with a stirrer to completely dissolve the hypromellose, thereby preparing a 0.5% hypromellose aqueous solution. . The benzoic acid salt of Compound A (in the Examples section, the benzoic acid salt of Compound A was used. Therefore, hereinafter “Compound A” refers to the benzoic acid salt of Compound A) was ground in an agate mortar and added to 0.5% hypromellose. It was suspended in an aqueous solution to a predetermined concentration, and ultrasonic treatment was performed to obtain a homogeneous suspension. The suspension containing Compound A was administered orally once daily for 6 consecutive days at a dose of 25 mg/kg/day. Dosages are expressed relative to the free form of Compound A.

Anti-mouse PD-1 antibody (anti-mPD-1 Ab) was incubated with anti-PD-1, CD279 (PD-1) monoclonal antibody (clone: RMP1-14, Thermo Fisher Scientific) immediately before administration. )) was prepared by diluting it with Dulbecco's phosphate buffered saline (nacalai tesque, INC.) to a predetermined concentration. On the first day of administration (day 1), the diluted product was administered intraperitoneally at a dose of 0.05 mg/body weight of anti-mouse PD-1 antibody.

To detect the expression of immune-related factors, flow cytometric analysis was performed for immunological monitoring. On day 7, tumors were sampled and tumor infiltrating lymphocytes were prepared using the Tumor Dissociation Kit, Mouse (Miltenyi Biotec). Thereafter, the isolated lymphocytes were stained with the following antibodies and then analyzed using a flow cytometer FACSVerse (BD Bioscience). For staining, CD45 monoclonal antibody (30-F11) eFluor 450, eBioscience ^TM (Thermo Fisher Scientific), Brilliant Violet 510 ^TM anti-mouse CD90.2 antibody (30-H2) (Bioscience) BioLegend), CD11b monoclonal antibody (M1/70) PE, Bioscience ^TM (Thermo Fisher Scientific), Ly-6G antibody anti-mouse (REA526), REAfinity ^TM (Miltenyi) Biotech) and CD8a monoclonal antibody (53-6.7) FITC, eBioscience ^TM (Thermo Fisher Scientific) were used.

The results are shown in Figures 1A and 1B.

On day 7, the ratio of gMDSCs (CD45-positive, CD90.2-negative, CD11b-positive, Ly6G-positive cells) in leukocytes (CD45-positive cells) of each group was analyzed by Dunnett's test, and the results showed that the group administered Compound A On day 7, the ratio of gMDSCs in leukocytes was significantly lower compared to the control group. On day 7, the ratio of CD8-positive cells (CD45-positive cells, CD90.2-positive cells and CD8-positive cells) in leukocytes (CD45-positive cells) of each group was analyzed by Dunnett's test, and the results were compared with Compound A and anti- The group treated with the combination of mouse PD-1 antibodies showed a significantly higher ratio of CD8 positive cells in leukocytes compared to the control group on day 7.

The results indicated that Compound A exhibited immunostimulatory activity.

Example 2: Effect of Biphenyl Compounds in Colon Cancer Cell Transplantation Model

MC38 cell suspension was inoculated into the right side of 6-week-old C57BL/6JJcl mice (Clea Japan, Inc.) or CB17/Icr-Prkdc[scid]/CrlCrlj (Charles River Laboratories Japan Inc.). It was implanted subcutaneously in the chest at 2.0 x 10 ⁶ cells/0.1 ml. CB17/Icr-Prkdc[scid]/CrlCrlj mice are mice with immunodeficiency due to lack of T-cells and B-cells in the peripheral blood.

Tumors were grown until they reached a TV of 50 to 300 mm ³ after implantation. Digimatic calipers were used to measure tumor diameter. The major and minor diameters of the tumor were measured, and TV was calculated from the equation below.

TV (mm ³ ) = Major diameter (mm) x Minor diameter (mm) x Minor diameter (mm)/2

By a stratified random assignment method using TV as an indicator, 10 C57BL/6JJcl mice were assigned to each group, and 5 CB17/Icr-Prkdc[scid]/CrlCrlj mice were assigned to each group. The day on which grouping was performed was defined as day 0.

An electronic animal scale was used to measure body weight. The body weight change ratio (BWCn) on the nth day was calculated from the body weight on the nth day (BWn) according to the formula below.

Weight change ratio BWCn (%) = (BWn-BW0)/BW0 x 100

To adjust the hypromellose concentration to 0.5 w/v%, an appropriate amount of water for injection as described in the Japanese Pharmacopoeia was added, and then the mixture was stirred with a stirrer to completely dissolve the hypromellose, thereby preparing a 0.5% hypromellose aqueous solution. . The benzoic acid salt of Compound A was ground in an agate mortar, then suspended in 0.5% hypromellose aqueous solution to a predetermined concentration, and ultrasonic treatment was performed to obtain a homogeneous suspension. The suspension containing Compound A was administered orally once daily for 21 consecutive days at a dose of 25 mg/kg/day. Dosages are expressed relative to the free form of Compound A.

The results are shown in Figures 2A and 2B and Tables 1 and 2.

[Table 1]

[Table 2]

On day 22, the TV of each group was analyzed by Aspin-Welch t-test, and the results showed an anti-tumor effect in the wild-type mouse model, with the Compound A administered group having a significantly lower TV compared to the control group. indicated that Meanwhile, the Compound A administered group in the immunodeficient mouse model did not show any antitumor effect compared to the control group.

Example 3: Effect of a combination of biphenyl compounds and immune checkpoint molecular modulators in a colon cancer cell transplantation model

MC38 cell suspension was implanted subcutaneously at 2.0 x 10 ⁶ cells/0.1 ml into the right thorax of 6-week-old C57BL/6JJcl mice (Clea Japan, Inc.).

Tumors were grown until they reached a TV of 50 to 300 mm ³ after implantation. Digimatic calipers were used to measure tumor diameter. The major and minor diameters of the tumor were measured, and TV was calculated from the equation below.

TV (mm ³ ) = Major diameter (mm) x Minor diameter (mm) x Minor diameter (mm)/2

By a stratified random assignment method using TV as an indicator, 10 mice were assigned to each group. The day on which grouping was performed was defined as day 0.

An electronic animal scale was used to measure body weight. The body weight change ratio (BWCn) on the nth day was calculated from the body weight on the nth day (BWn) according to the formula below.

Weight change ratio BWCn (%) = (BWn-BW0)/BW0 x 100

To adjust the hypromellose concentration to 0.5 w/v%, an appropriate amount of water for injection as described in the Japanese Pharmacopoeia was added, and then the mixture was stirred with a stirrer to completely dissolve the hypromellose, thereby preparing a 0.5% hypromellose aqueous solution. . The benzoic acid salt of Compound A was ground in an agate mortar, then suspended in 0.5% hypromellose aqueous solution to a predetermined concentration, and ultrasonic treatment was performed to obtain a homogeneous suspension. The suspension containing Compound A was administered orally once daily for 21 consecutive days at a dose of 25 mg/kg/day. Dosages are expressed relative to the free form of Compound A.

Anti-mouse PD-1 antibody (anti-mPD-1 Ab) was prepared by adding anti-PD-1, CD279 (PD-1) monoclonal antibody (clone: RMP1-14, Bio It was prepared by diluting to a predetermined concentration with Otsuka Pharmaceutical Factory, Inc.). On the first day of administration (day 1), the diluted product was administered intraperitoneally at a dose of 0.05 mg/body weight of anti-mouse PD-1 antibody.

The results are shown in Figure 3 and Table 3.

[Table 3]

On day 22, the TV of each group was analyzed by Aspin-Welch t-test, and the results were obtained for the Compound A group or the single-agent group administered anti-mouse PD-1 antibody and Compound A + anti-mouse PD. It was shown that both combination administration groups administered -1 antibody exhibited anti-tumor effect with significantly lower TV compared to the control group. Additionally, the combination group administered Compound A + anti-mouse PD-1 antibody had significantly lower TV and higher anti-mouse group compared to the single-agent group administered Compound A or anti-mouse PD-1 antibody. showed antitumor effect.

The mean weight change ratio of the combination administration group did not show any enhanced toxicity compared to the single-agent group administered Compound A or anti-mouse PD-1 antibody alone.

Claims (26)

4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-) for use in the treatment of a subject having cancer. An antitumor agent comprising 2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof, wherein the treatment includes administering an immune checkpoint molecule modulator to the subject. . The anti-tumor agent according to claim 1, wherein the immune checkpoint molecule modulator is at least one selected from the group consisting of a PD-1 pathway antagonist and a CTLA-4 pathway antagonist. The anti-tumor agent according to claim 1 or 2, wherein the immune checkpoint molecule modulator is a PD-1 pathway antagonist. The anti-tumor agent according to claim 2 or 3, wherein the PD-1 pathway antagonist is at least one selected from the group consisting of anti-PD-1 antibody, anti-PD-L1 antibody, and anti-PD-L2 antibody. The anti-tumor agent according to claim 2 or 3, wherein the PD-1 pathway antagonist is an anti-PD-1 antibody or an anti-PD-L1 antibody. The anti-tumor agent according to claim 2 or 3, wherein the PD-1 pathway antagonist is an anti-PD-1 antibody. The anti-tumor agent of claim 2, wherein the CTLA-4 pathway antagonist is an anti-CTLA-4 antibody. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl] as active ingredient An anti-tumor effect enhancer for an immune checkpoint molecular modulator, comprising phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]- An antitumor agent for treating cancer patients given an immune checkpoint molecule modulator, comprising 2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-), containing immune checkpoint molecular modulators An antitumor agent for treating cancer patients given methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro- for use in treating tumors by administration in combination with immune checkpoint molecule modulators 4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-( for use in enhancing the antitumor effect of immune checkpoint molecular modulators 2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4 for use in treating tumors in cancer patients given immune checkpoint molecule modulators -(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]- An immune checkpoint molecular modulator for use in treating tumors in cancer patients given 2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl) for use in treating tumors -Propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and a combination of an immune checkpoint molecular modulator. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4- for preparing antineoplastic agents administered in combination with immune checkpoint molecular modulators. Use of (2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluo for preparing anti-tumor effect enhancer to enhance the anti-tumor effect of immune checkpoint molecular modulator. Use of rho-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro for preparing antineoplastic agents for treating cancer patients given immune checkpoint molecular modulators Use of -4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]- Use of an immune checkpoint molecular modulator to prepare an anti-tumor agent for the treatment of cancer patients given 2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-) for preparing antitumor agents Propyl) phenyl] phenyl] -2-fluoro-benzonitrile or a salt thereof and use of an immune checkpoint molecule modulator. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-) to a subject in need of treatment for a tumor. A method of treating a tumor comprising administering a combination of methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and an immune checkpoint molecule modulator. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4- for subjects in need of enhancement of the anti-tumor effect of an immune checkpoint molecular modulator. A method for enhancing the antitumor effect of an immune checkpoint molecular modulator, comprising administering (2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-) to a subject in need of treatment for a tumor. A method of treating a tumor in a cancer patient given an immune checkpoint molecule modulator, comprising administering methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluo, comprising administering an immune checkpoint molecule modulator to a subject in need of treatment of a tumor. A method of treating a tumor in a cancer patient given rho-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-) to a subject in need of treatment for a tumor. A method of treating a tumor comprising administering a combination of methyl-propyl)phenyl]phenyl]-2-fluoro-benzonitrile or a salt thereof and an immune checkpoint molecular modulator. 4-[5-[(3S)-3-aminopyrrolidine-1-carbonyl]-2-[2-fluoro-4-(2-hydroxy-2-methyl-propyl)phenyl]phenyl]- A pharmaceutical composition for preventing and/or treating tumors, comprising 2-fluoro-benzonitrile or a salt thereof and an immune checkpoint molecule modulator.

Images